• Korean Journal of Microbiology
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• v.47 no.3
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• pp.194-199
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• 2011

An alkalophilic bacterium producing alkaline protease was isolated from waste water and solar saltern sample and identified as Bacillus pseudofirmus HS-54 based on morphological, biochemical characteristics as well as 16S-rRNA gene sequencing. The HS-54 protease was purified to homogeneity using ammonium sulfate precipitation, DEAE cellulose column chromatography, and sephadex G-100 gel filtration with a 4.0 purification fold. The molecular mass of the purified enzyme was estimated by SDS-PAGE to be 27 kDa. The optimal pH and temperature for the purified protease activity were 10.0 and $50^{\circ}C$, respectively. The purified enzyme was relatively stable at the pH range of 6.0-11.0 and at the temperature below $50^{\circ}C$. This enzyme was activated by $Ca^{2+}$ and $Mg^{2+}$ and inhibited by $Hg^{2+}$, $Cu^{2+}$, $Zn^{2+}$, $Al^{3+}$, $Ag^{2+}$. And this enzyme was strongly inhibited by PMSF, suggesting that it belongs to the serine protease superfamily.

• Microbiology and Biotechnology Letters
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• v.32 no.3
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• pp.230-237
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• 2004

In this study nitroreductase from Pseudomonas sp. HK-6 capable of degrading 2,4,6-trinitrotoluene (TNT) was characterized. Through a series of purification process including ammonium sulfate precipitation, DEAE-sepharose, and Q-sepharose, three different fractions I, II, and III having the enzyme activity of NTRs whose molecular weights were approximately 27 kDa were detected in fractions from HK-6 cells. Specific activity of the three fractions were approximately 4.85 unit/mg, 5.47 unit/mg, and 5.01 unit/mg, and concentrated to 9.0-, 10.1-, and 9.3-fold compared to crude extract, respectively. The optimal pH and temperature for the three NTR fractions were approximately 7.5 and $30^{\circ}C$, respectively. Metal ions, $Ag^{＋}$ , $Cu^{ 2+}$, $Hg^{2＋}$ inhibited approximately 70% of enzymes activities of all NTR, while $Fe^{2＋}$ did not stimulate or inhibit the activities. Monitoring the effect of chemicals on the enzyme activity revealed that those NTR fractions lost enzyme activity in presence of $\beta$-mercaptoethanol, but were a little influenced by dithiothreitol, EDTA and NaCl. The three NTR fractions demonstrated enzyme activities for nitrobenzene and RDX as well as TNT.

• Economic and Environmental Geology
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• v.52 no.5
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• pp.323-336
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• 2019

The Mesozoic activity on the Korean Peninsula is mainly represented by the Triassic post-collisional, Jurassic orogenic, and Cretaceous post-orogenic igneous activities. The diversity of mineralization by each geological period came from various geothermal systems derived from the geochemical characteristics of magma with different emplacement depth. The Cretaceous metallic mineralization has been carried out over a wide range of time periods from ca. 115 to 45 Ma (main stage; ca. 100 to 60 Ma) related to post-orogenic igneous activity, and spatial distribution patterns of most metal deposits are concentrated along small granitic stocks. The late Cretaceous metal deposits in the Gyeonggi and Yeongnam massifs are generally distributed along the boundary among the Gongju-Eumseong fault system and the Yeongdong-Gwangju fault system and the Gyeongsang Basin, most of them are in the form of a distal epithermal~mesothermal Au-Ag vein or a transitional mesothermal Zn-Pb-Cu vein. On the other hand, diverse metal commodities in the Taebaeg Basin, the Okcheon metamorphic belt and the Gyeongsang Basin are produced from various deposit types such as skarn, carbonate-replacement, vein, porphyry, breccia pipe, and Carlin type. In the late Cretaceous metallic mineralization, various mineral deposits and commodities were induced not only by the pathway of the hydrothermal solution, but also by the diversity of precipitation environment in the proximity difference of the granitic rocks. The diversity of these types of Cretaceous deposits is fundamentally dependent on the geochemical characteristics such as degree of differentiation and oxidation state of related igneous rocks, and ore-forming fluids generally exhibit the evolutionary characteristics of intermediate- to low-sulfur hydrothermal fluids.

• Economic and Environmental Geology
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• v.54 no.1
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• pp.49-60
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• 2021

The Geumhwa Au-Ag deposit is located within the Cretaceous Gyeongsang basin. Mineral paragenesis can be divided into two stages (stage I and II) by major tectonic fracturing. Stage II is economically barren. Stage I, at which the precipitation of major ore minerals occurred, is further divided into three substages(early, middle and late) with paragenetic time based on minor fractures and discernible mineral assemblages: early substage, marked by deposition of pyrite with minor wolframite; middle substage, characterized by introduction of electrum and base-metal sulfides with Cu-As and/or Cu-Sb sulfosalts; late substage, marked by hematite and Bi-sulfosalts with secondary minerals. Changes in vein mineralogy reflect decreases in temperature and sulfur fugacity with a concomitant increase in oxygen fugacity. Fluid inclusion data indicate progressive decreases in temperature and salinity within each substage with increasing paragenetic time. During the early portion of stage I, high-temperature (≥410℃), high-salinity fluids (up to ≈44 equiv. wt. % NaCl) formed by condensation during decompression of a magmatic vapor phase. During waning of early substage, high-temperature, high-salinity fluids gave way to progressively cooler, more dilute fluids associated with main Au-Ag mineralization (middle) and finally to ≈180℃ and ≥0.7 equiv. wt. % NaCl fluids associated with hematite and sulfosalts (± secondary) mineralization (late substage). These trends are interpreted to indicate progressive mixing of high- and medium to low-salinity hydrothermal fluids with cooler, more dilute, oxidizing meteoric waters. The Geumhwa Au-Ag deposit may represent a vein-type system transitional between porphyry-type and epithermal-type.

• Journal of Applied Biological Chemistry
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• v.44 no.3
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• pp.113-118
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• 2001

Xylose (glucose) isomerase was purified to homogeneity from cell-extracts of Streptomyces chibaensis J-59 via ammonium sulfate precipitation followed by chromatography on DEAE-cellulose, and gel filtration on Sephacryl S-300. The purified enzyme is a homotetramer with a native molecular mass of 180 kDa and a subunit molecular mass of 44 kDa. The amino acid N-terminal sequence of glucose isomerase from S. chibaensis J-59 was determined to be Ser-Tyr-Gln-Pro-Thr-Pro-Glu-Asp-Arg-Phe-Thr-Phe-Gly-Leu. The first 14 amino acids of the N-terminal sequence of the enzyme showed strong analogies with N-terminal sequences of glucose isomerase produced by other Streptomyces spp. The optimum pH and temperature for activity were 7.5 and 85, respectively. The purified enzyme required $Mg^{2+}$, $Co^{2+}$, and $Mn^{2+}$ for the activity, $Mg^{2+}$ being the most effective. The enzyme was not inhibited by $Ca^{2+}$, but was inhibited by $Hg^{2+}$, $Ag^+$, and $Cu^{2+}$. The $K_m$, $V_{max}$, and $k_{cat}$ values of S. chibaensis J-59 isomerase for glucose were 83 mM, 40.9 U/mg, and $1,843min^{-1}$, respectively. In the presence of $Co^{2+}$, cell-free enzymes retained 100% without loss of activities by the heat-treatment at $70^{\circ}C$ for 7 days. The enzyme retained 50% residual activity after heating at $85^{\circ}C$ for 13.5 h, at $90^{\circ}C$ for 126 min. The enzyme is more thermostable than any other glucose isomerases of Streptomyces spp.

• Journal of Microbiology
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• v.35 no.2
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• pp.109-116
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• 1997

An extracellular proteinase of Candida albicans was purified by a combination of 0~75% ammonium sulfate precipitation, DEAE Sepharose Fast Flow ion exchange chromatography, and Sephacryl S-200 HR molecular sieve chromatography. Its mlecular weight was approximately 41 kDa on SDS-PAGE and isoelectric point was 4.4. The enzyme was inhibited by pepstain A. Optimum enzyme activity ranged from pH 2.0 to 3.5 with its maximum at pH 2.5 and a temperature of 45$^{\circ}C$. The addition of divalent cations, $Ca^{2+}$, Zn$^{2+}$ and $Mg^{2+}$, resulted in no significant inhibition of enzymatic activity. However, some inhibitory effects were observed by Fe$^{2+}$, Ag$^{2+}$ and Cu$^{2+}$. With BSA as substrate, an apparent $K_m$ was determined to be 7$\times$10$^{-7}$ M and $K_i$, using pepstatin A as an inhibitor, was 8.05$\times$10$^{-8}$ M. N-terminal amino acid sequence was QAVPVTLXNEQ. Degradation of BSA and fibronectin was shown but not collagen, hemoglobin, immunoglobulin G, or lysozyme. The enzyme preferred peptides with Glu and Leu at the P$_1$ position, but the enzyme activity was highly reduced when the P$_2$ position was phe or pro. This enzyme showed antigenicity against sera of patients with candidiasis.

• Economic and Environmental Geology
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• v.25 no.3
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• pp.233-243
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• 1992

Ore mineralization of the Hwanghak copper deposit in the Ogsan area occurred in three stages of quartz (stage I and II) and calcite (stage III) veining along fissures in Early Cretaceous sedimentary rocks. Ore minerals are pyrite, pyrrhotite, chalcopyrite (dominant), sphalerite, hematite, galena, and Ag-, Pb-, and Bi-sulfosalts. These were deposited during the first stage at temperatures between $370^{\circ}C$ and < $200^{\circ}C$ from fluids with salinities between 0.5 and 7.6 equiv. wt. % NaCl. There is evidence of boiling and this suggests pressures of less than 180 bars during the first stage. Equilibrium thermodynamic interpretation accompanying with mineral paragenesis and fluid inclusion data indicates that copper precipitation in the hydrothermal system occurred due to cooling and changing in chemical conditions ($fs_2$, $fo_2$, pH). Gradual temperature decrease from $350^{\circ}$ to $250^{\circ}C$ of ore fluids by boiling and mixing with less-evolved meteoric waters mainly led to copper deposition through destabilization of copper chloride complexes. Sulfur isotope values of sulfide minerals decrease systematically with paragenetic time from calculated ${\delta}^{34}S_{H_2S}$ values of 8.2 to 4.7‰. These values, together with the observed change from sulfide-only to sulfide-hematite assemblages and fluid inclusion data, suggest progressively more oxidizing conditions, with a corresponding increase of the $sulfate/H_2S$ ratio of hydrothermal fluids. Measured and calculated hydrogen and oxygen isotope valutls of ore-forming fluids suggest meteoric water dominance, approaching unexchanged meteoric water values.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.275-281
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• 2004

Our previous research has demonstrated that the bacterium, Pseudomonas sp. NFQ-l capable of utilizing quin­oline (2,3-benzopyridine) as the sole source of carbon, nitrogen, and energy was isolated and characterized [Yoon et ai. (2003) Kor. J. Biotechnol. Bioeng. 18(3):174-179]. In this study, we have found that Pseudomonas sp. NFQ-l could degrade quinoline as well as benzoate, and extended this work to characterize the catechol 1,2­dioxygenase (C1,2O) purified from the bacterium cultured in benzoate media. Initially, C1,2O has been purified by ammonium sulfate precipitation, gel permeation chromatography, and Source 15Q. After Source 15Q, puri­fication fold was increased to approximately 14.21 unit/mg. Molecular weight of C1,2O was about 33 kDa. Physicochemical characteristics (e.g., substrate specificity, Km, Vmax, pH, temperature and effect of inhibitors) of purified C1,2O were examined. C1,2O demonstrated the activity for catechol, 4-methylcatechol and 3-meth­ylcatechol as a substrate, respectively. The Km and Vmax value of C1,2O for catechol was 38.54 ${\mu}M$ and $25.10\;{\mu}mol{\cdot}min^{-1}{\cdot}mg^{-1}.$ The optimal temperature of C1,2O was $30^{\circ}C$ and the optimal pH was approximately 8.5. Metal ions such as $Ag^+,\;Hg^+,\;Ca^{2+},\;and\;Cu^{2+}$ show the inhibitory effect on the activity of C1,2O. N-terminal amino sequence of C1,2O was analyzed as ^1TVKISQSASIQKFFEEA^{17}.$ In this work, we found that the amino acid sequence of NFQ-l showed the sequence homology of 82, 71, 59 and $53\%$ compared with C1,2O from Pseudomonas aeruginosa PA0l, Pseudomonas arvilla C-1., P. putida KT2440 and Pseudomonas sp. CA10, respectively.

• Economic and Environmental Geology
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• v.37 no.1
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• pp.1-19
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• 2004

Metallic deposits in Korea have a variety of genetic types such as hydrothermal veins, skarns, hydrothermal replacement and alaskite deposits and so on. Geological, mineralogical and geochemical features including host rock, wall-rock alteration, ore and gangue mineralogy, mineral texture and secondary mineralogy related to weathering process control the environmental signatures of mining areas. The environmental signatures of metallic deposits closed from early 1970s to late 1990s in Korea show complicate geochemistry and mineralogy due to step weathering of primary and secondary minerals such as oxidation-precipitation-remobilization. The potentiality of low pH and high heavy metal Concentration s from acid mine drainage is great in base-metal deposits associated with polymetallic mineralization, breccia-pipe type and Cretaceous hydrothermal Au veins with the amount of pyrite whereas skam, hydrothermal replacement, hydrothermal Cu and Au-Ag vein deposits are in low contamination possibility. The geoenvironmental models reflecting the various geologic features closely relate to disuibution of sulfides and carbonates and their ratios and finally effect on characteristics of environmental signatures such as heavy metal species and their concentrations in acid mine drainage.